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Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition

View Article: PubMed Central - PubMed

ABSTRACT

Although increases in cardiovascular load (pressure overload) are known to elicit ventricular remodeling including cardiomyocyte hypertrophy and interstitial fibrosis, the molecular mechanisms of pressure overload or AngII -induced cardiac interstitial fibrosis remain elusive. In this study, serpinE2/protease nexin-1 was over-expressed in a cardiac fibrosis model induced by pressure-overloaded via transverse aortic constriction (TAC) in mouse. Knockdown of serpinE2 attenuates cardiac fibrosis in a mouse model of TAC. At meantime, the results showed that serpinE2 significantly were increased with collagen accumulations induced by AngII or TGF-β stimulation in vitro. Intriguingly, extracellular collagen in myocardial fibroblast was reduced by knockdown of serpinE2 compared with the control in vitro. In stark contrast, the addition of exogenous PN-1 up-regulated the content of collagen in myocardial fibroblast. The MEK1/2- ERK1/2 signaling probably promoted the expression of serpinE2 via transcription factors Elk1 in myocardial fibroblast. In conclusion, stress-induced the ERK1/2 signaling pathway activation up-regulated serpinE2 expression, consequently led accumulation of collagen protein, and contributed to cardiac fibrosis.

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Related in: MedlinePlus

The expression of SerpinE2 increases in transverse aortic constriction (TAC)-induced mouse cardiac myocardial fibrosis model.(a) Collagen I/III (col1a/col3a) mRNA levels was detected by qRT-PCR. n = 8. *P < 0.05 vs. control. (b) Sircol™ Collagen Assay was used to quantify the total collagen contents in myocardial tissue from TAC models. n = 10. *P < 0.05 vs. control. (c) The expression of serpinE2 mRNA in plasma (n = 5, *P < 0.05 vs. control) and myocardium (n = 8, *P < 0.05 vs. control) of TAC mice using qRT-PCR. (d) The level of serpinE2 protein in plasma (n = 7, *P < 0.05 vs. control) and myocardium (n = 10, *P < 0.01 vs. control) of TAC mice using ELISA. (e) Western blot analysis of serpinE2 protein and statistical analysis (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 1. (f) Representative sections of heart with Masson staining. The fibrotic tissues are stained as blue, as indicated by the arrows. Scale bars: 200 μm. (g) Collagen deposition was quantified by automated image analysis and expressed as percentage of tissue area. n = 6. (h) The relative protein level of serpinE2 and collagen was detected by ELISA and Sircol™ Collagen Assay in Knockdown -serpinE2 mice. n = 6. (i) The protein of collagen, serpinE2 and α-sma were decreased in Knockdown-serpinE2 mice. Full-length blots/gels are presented in Supplementary Figure 2 and Supplementary Figure 3 (n = 6, #P < 0.05 vs. TAC + LV-NC).
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f1: The expression of SerpinE2 increases in transverse aortic constriction (TAC)-induced mouse cardiac myocardial fibrosis model.(a) Collagen I/III (col1a/col3a) mRNA levels was detected by qRT-PCR. n = 8. *P < 0.05 vs. control. (b) Sircol™ Collagen Assay was used to quantify the total collagen contents in myocardial tissue from TAC models. n = 10. *P < 0.05 vs. control. (c) The expression of serpinE2 mRNA in plasma (n = 5, *P < 0.05 vs. control) and myocardium (n = 8, *P < 0.05 vs. control) of TAC mice using qRT-PCR. (d) The level of serpinE2 protein in plasma (n = 7, *P < 0.05 vs. control) and myocardium (n = 10, *P < 0.01 vs. control) of TAC mice using ELISA. (e) Western blot analysis of serpinE2 protein and statistical analysis (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 1. (f) Representative sections of heart with Masson staining. The fibrotic tissues are stained as blue, as indicated by the arrows. Scale bars: 200 μm. (g) Collagen deposition was quantified by automated image analysis and expressed as percentage of tissue area. n = 6. (h) The relative protein level of serpinE2 and collagen was detected by ELISA and Sircol™ Collagen Assay in Knockdown -serpinE2 mice. n = 6. (i) The protein of collagen, serpinE2 and α-sma were decreased in Knockdown-serpinE2 mice. Full-length blots/gels are presented in Supplementary Figure 2 and Supplementary Figure 3 (n = 6, #P < 0.05 vs. TAC + LV-NC).

Mentions: It has well been accepted that surgical transverse aortic constriction (TAC) induces cardiac fibrosis after 4 weeks1718. As shown in Fig. 1a,b,f, the marked collagen deposition were observed in TAC group, suggesting dramatic fibrosis along with pathphysiological process of cardiac hypertrophy in TAC mice at 4 weeks. Approximately 85% of total myocardial collagen is type I, and 11% is collagen type III in the heart, which typically form thin fibers and maintain the elasticity of the matrix network319 in the physiological condition. Therefore, we have a reason to believe that TAC would cause cardiac fibrosis. To test this hypothesis, Collagen types I (col1a) and III mRNA (col3a) were evaluated by RT-PCR. The expression of col1a and col3a mRNA were increased in the interstitial tissue of TAC mouse (Fig. 1a). As expected, total collagen content was increased significantly by about 1.30 fold detected by Sircol™ Collagen Assay Kit (Fig. 1b).


Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition
The expression of SerpinE2 increases in transverse aortic constriction (TAC)-induced mouse cardiac myocardial fibrosis model.(a) Collagen I/III (col1a/col3a) mRNA levels was detected by qRT-PCR. n = 8. *P < 0.05 vs. control. (b) Sircol™ Collagen Assay was used to quantify the total collagen contents in myocardial tissue from TAC models. n = 10. *P < 0.05 vs. control. (c) The expression of serpinE2 mRNA in plasma (n = 5, *P < 0.05 vs. control) and myocardium (n = 8, *P < 0.05 vs. control) of TAC mice using qRT-PCR. (d) The level of serpinE2 protein in plasma (n = 7, *P < 0.05 vs. control) and myocardium (n = 10, *P < 0.01 vs. control) of TAC mice using ELISA. (e) Western blot analysis of serpinE2 protein and statistical analysis (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 1. (f) Representative sections of heart with Masson staining. The fibrotic tissues are stained as blue, as indicated by the arrows. Scale bars: 200 μm. (g) Collagen deposition was quantified by automated image analysis and expressed as percentage of tissue area. n = 6. (h) The relative protein level of serpinE2 and collagen was detected by ELISA and Sircol™ Collagen Assay in Knockdown -serpinE2 mice. n = 6. (i) The protein of collagen, serpinE2 and α-sma were decreased in Knockdown-serpinE2 mice. Full-length blots/gels are presented in Supplementary Figure 2 and Supplementary Figure 3 (n = 6, #P < 0.05 vs. TAC + LV-NC).
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f1: The expression of SerpinE2 increases in transverse aortic constriction (TAC)-induced mouse cardiac myocardial fibrosis model.(a) Collagen I/III (col1a/col3a) mRNA levels was detected by qRT-PCR. n = 8. *P < 0.05 vs. control. (b) Sircol™ Collagen Assay was used to quantify the total collagen contents in myocardial tissue from TAC models. n = 10. *P < 0.05 vs. control. (c) The expression of serpinE2 mRNA in plasma (n = 5, *P < 0.05 vs. control) and myocardium (n = 8, *P < 0.05 vs. control) of TAC mice using qRT-PCR. (d) The level of serpinE2 protein in plasma (n = 7, *P < 0.05 vs. control) and myocardium (n = 10, *P < 0.01 vs. control) of TAC mice using ELISA. (e) Western blot analysis of serpinE2 protein and statistical analysis (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 1. (f) Representative sections of heart with Masson staining. The fibrotic tissues are stained as blue, as indicated by the arrows. Scale bars: 200 μm. (g) Collagen deposition was quantified by automated image analysis and expressed as percentage of tissue area. n = 6. (h) The relative protein level of serpinE2 and collagen was detected by ELISA and Sircol™ Collagen Assay in Knockdown -serpinE2 mice. n = 6. (i) The protein of collagen, serpinE2 and α-sma were decreased in Knockdown-serpinE2 mice. Full-length blots/gels are presented in Supplementary Figure 2 and Supplementary Figure 3 (n = 6, #P < 0.05 vs. TAC + LV-NC).
Mentions: It has well been accepted that surgical transverse aortic constriction (TAC) induces cardiac fibrosis after 4 weeks1718. As shown in Fig. 1a,b,f, the marked collagen deposition were observed in TAC group, suggesting dramatic fibrosis along with pathphysiological process of cardiac hypertrophy in TAC mice at 4 weeks. Approximately 85% of total myocardial collagen is type I, and 11% is collagen type III in the heart, which typically form thin fibers and maintain the elasticity of the matrix network319 in the physiological condition. Therefore, we have a reason to believe that TAC would cause cardiac fibrosis. To test this hypothesis, Collagen types I (col1a) and III mRNA (col3a) were evaluated by RT-PCR. The expression of col1a and col3a mRNA were increased in the interstitial tissue of TAC mouse (Fig. 1a). As expected, total collagen content was increased significantly by about 1.30 fold detected by Sircol™ Collagen Assay Kit (Fig. 1b).

View Article: PubMed Central - PubMed

ABSTRACT

Although increases in cardiovascular load (pressure overload) are known to elicit ventricular remodeling including cardiomyocyte hypertrophy and interstitial fibrosis, the molecular mechanisms of pressure overload or AngII -induced cardiac interstitial fibrosis remain elusive. In this study, serpinE2/protease nexin-1 was over-expressed in a cardiac fibrosis model induced by pressure-overloaded via transverse aortic constriction (TAC) in mouse. Knockdown of serpinE2 attenuates cardiac fibrosis in a mouse model of TAC. At meantime, the results showed that serpinE2 significantly were increased with collagen accumulations induced by AngII or TGF-&beta; stimulation in vitro. Intriguingly, extracellular collagen in myocardial fibroblast was reduced by knockdown of serpinE2 compared with the control in vitro. In stark contrast, the addition of exogenous PN-1 up-regulated the content of collagen in myocardial fibroblast. The MEK1/2- ERK1/2 signaling probably promoted the expression of serpinE2 via transcription factors Elk1 in myocardial fibroblast. In conclusion, stress-induced the ERK1/2 signaling pathway activation up-regulated serpinE2 expression, consequently led accumulation of collagen protein, and contributed to cardiac fibrosis.

No MeSH data available.


Related in: MedlinePlus